The term avermectin (previously referred to as C-076) is used to describe a series of compounds isolated from the fermentation broth of an avermectin producing strain of Streptomyces avermitilis and derivatives thereof. The morphological characteristics of the culture are completely described in U.S. Pat. No. 4,310,519. The avermectin compounds are a series of macrolides, each of which is substituted at the 13 position with a 4-(alpha-L-oleandrosyl)-alpha-L-oleandrose group. The avermectin compounds and the instant derivatives thereof have a very high degree of anthelmintic and anti-parasitic activity.
The avermectin series of compounds isolated from the fermentation broth have the following structure: ##STR1## wherein R.sub.4 is the 4'-.alpha.-(L-oleandrosyl)-.alpha.-L-oleandrosyloxygroup of the structure ##STR2## and wherein A at the 22,23 position indicates a single or a double bond;
R.sub.1 is a hydrogen or hydroxy and is present only when A indicates a single bond; PA0 R.sub.2 is iso-propyl or sec-butyl; and PA0 R.sub.3 is methoxy or hydroxy. PA0 3',3"-Bisdesmethylavermectin B1a and B1b PA0 3',3"-Bisdesmethylavermectin B2a and B2b PA0 3"-Desmethylavermectin B1a and B1b PA0 3"-Desmethylavermectin B2a and B2b PA0 3',3"-Bisdesmethyl-25-cyclohexyl-25-de-(2-butyl)avermectin B2a PA0 3',3"-Bisdesmethyl-25-cyclopentyl-25-de-(2-butyl)avermectin B2a PA0 3',3"-Bisdesmethyl-25-(3-thienyl)-25-de-(2-butyl)avermectin B2a PA0 3',3"-Bisdesmethyl-25-(3furyl)-25-de-(2-butyl)avermectin B2a PA0 3',3"-Bisdesmethyl-25-(1-methylthioethyl)-25-de-(2-butyl)avermectin B1a. PA0 and wherein R.sub.1 is hydrogen or hydroxy or oxo, or A represents a double bond and R.sub.1 is absent; PA0 R.sub.2 is methyl, ethyl, an alpha-branched C.sub.3 -C.sub.8 alkyl, alkenyl, alkynyl, alkoxyalkyl or alkylthioalkyl group; a C.sub.3 -C.sub.8 cycloalkyl or C.sub.5 -C.sub.8 cycloalkenyl group, either of which may optionally be substituted by methylene or one or more C.sub.1 -C.sub.4 alkyl groups or halo atoms; or a 3 to 6 membered oxygen or sulfur containing heterocyclic ring which may be saturated, or fully or partly unsaturated and which may optionally be substituted by one or more C.sub.1 -C.sub.4 alkyl groups or halo atoms; PA0 R.sub.3 is hydroxy, loweralkoxy, loweralkanoyloxy, oxo or oxime; PA0 R.sub.4 is ##STR4## where R.sub.5 is NR.sub.6 R.sub.7, R.sub.6 is substituted loweralkanoyl, where the substituent is halogen, hydroxy, loweralkoxy, phenoxy, loweralkylthio, loweralkylsulfinyl, loweralkylsulfonyl, amino, loweralkanoylamino, loweralkylamino, haloloweralkoxycarbonylamino, oxo, carboxy or loweralkoxycarbonyl; or R.sub.6 is cycloloweralkanoyl, or benzoyl, or substituted benzoyl, wherein the substituent is halogen, loweralkoxy, sulfonamido, amino, loweralkylamino, diloweralkylamino or loweralkanoylamino; or R.sub.6 is nicotinoyl; PA0 R.sub.7 is hydrogen, loweralkyl, substituted loweralkyl where the substituent is phenyl, hydroxy, loweralkoxy, amino, loweralkylamino, loweralkanoylamino, methylthio, methylsulfonyl or methylsulfinyl; PA0 or R.sub.5 = ##STR5## R.sub.5 =--NH--CO--NR.sub.8 R.sub.9, R.sub.8, R.sub.9 and R.sub.10 are independently hydrogen or loweralkyl; PA0 or R.sub.5 =--NH--CN. PA0 wherein R.sub.1 is hydrogen or hydroxy, or A represents a double bond and R.sub.1 is absent; PA0 R.sub.2 iso-propyl, sec-butyl, or an alpha-branched C.sub.3 -C.sub.8 alkenyl group; and PA0 R.sub.3 is hydroxy or oxime; PA0 R.sub.4 is ##STR6## where R.sub.5 is NR.sub.6 R.sub.7, R.sub.6 is substituted loweralkanoyl, where the substituent is halogen, hydroxy, loweralkoxy, phenoxy, loweralkylthio, loweralkylsulfinyl, loweralkylsulfonyl, amino, loweralkanoylamino, loweralkylamino, haloloweralkoxycarbonylamino, oxo, carboxy or alkoxycarbonyl; or R.sub.6 is cycloloweralkanoyl, or benzoyl, or substituted benzoyl, wherein the substituent is halogen, loweralkoxy, sulfonamido, amino, loweralkylamino, diloweralkylamino or loweralkanoyl; or R.sub.6 is nicotinoyl; PA0 R.sub.7 is hydrogen, loweralkyl, substituted loweralkyl where the substituent is phenyl, hydroxy, loweralkoxy, amino, loweralkanoylamino, loweralkanoyl, methylthio, methylsulfonyl or methylsulfinyl; PA0 or R.sub.5 =--NH--CO--NR.sub.8 R.sub.9, PA0 where R.sub.8 and R.sub.9 and R.sub.10 are independently hydrogen or loweralkyl. PA0 R.sub.1 is hydrogen or hydroxy, or A represents a double bond and R.sub.1 is absent; PA0 R.sub.2 is 2-propyl, 2-butyl, 2-buten-2-yl, 2-penten-2-yl, or 4-methyl-2-penten-2-yl; PA0 R.sub.3 is hydroxy; PA0 R.sub.4 is ##STR7## where R.sub.5 is NR.sub.6 R.sub.7, R.sub.6 is substituted loweralkanoyl, where the substituent is halogen, hydroxy, loweralkoxy, phenoxy, loweralkylthio, loweralkylsulfinyl, loweralkylsulfonyl, amino, loweralkanoyl, loweralkylamino, haloloweralkoxycarbonylamino, oxo, carboxy or loweralkoxycarbonyl; or R.sub.6 is lowercycloalkanoyl, or benzoyl, or substituted benzoyl, wherein the substituent is halogen, loweralkoxy, sulfonamido, amino, loweralkylamino, diloweralkylamino or loweralkanoyl; or R.sub.6 is nicotinoyl; PA0 R.sub.7 is hydrogen, loweralkyl, substituted loweralkyl where the substituent is phenyl; PA0 or R.sub.5 =--NH--CO--NR.sub.8 R.sub.9, PA0 where R.sub.8 and R.sub.9 and R.sub.10 are independently hydrogen or loweralkyl. PA0 4"-deoxy-4"-epi-methoxyacetylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-methoxyacetylaminoavermectin B1a/B1b PA0 4"-deoxy-22,23-dihydro-4"-epi-methoxyacetylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-methoxyacetylaminoavermectin B2a/B2b PA0 25-cylopentyl-25-de-(1-methylpropyl)-4"-epi-methoxyacetylaminoavermectin B1a PA0 4"-deoxy-4"-epi-(N-methoxyacetyl-N-methylamino)avermectin B1a/B1b PA0 4"-deoxy-4"-epi-methylthioacetylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-methylsulfinylacetylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-methylsulfonylacetylaminoavermectin B1a/B1b PA0 4'-deoxy-4'-epi-methylthioacetylaminoavermectin B1a/B1b monosaccharide PA0 4"-deoxy-4"-epi-glycylaminoavermectin B1a/B1b PA0 4"-epi-(N-acetylglycylamino)-4"-deoxyavermectin B1a/B1b PA0 4"-deoxy-4"-epi-(2-hydroxypropionylamino)avermectin B1a/B1b PA0 4"-deoxy-4"-epi-phenoxyacetylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-(2,2,2-trichloroethoxycarbonyl)-glycylaminoavermectin B1a/B1b PA0 4"-epi-benzoylamino-4"-deoxyavermectin B1a/B1b PA0 4"-epi-(4-chlorobenzoylamino)-4"-deoxyavermectin B1a/B1b PA0 4"-epi-(4-methoxybenzoylamino)-4"-deoxyavermectin B1a/B1b PA0 4"-epi-(3-chloro-4-aminosulfonylbenzoylamino)-4"-deoxyavermectin B1a/B1b PA0 4"-deoxy-4"-epi-nicotinoylamino-4"-avermectin B1a/B1b PA0 4"-epi-cyclohexylcarbonylamino-4"-deoxyavermectin B1a/B1b PA0 4"-epi-(3-chloro-4-dimethylaminobenzoylamino)-4"-deoxy avermectin B1a/B1b PA0 4"-deoxy-4"-epi-(2,2,2-trimethylacetylamino)-avermectin B1a/B1b PA0 4"-epi-chloroacetylamino-4"-deoxyavermectin B1a/B1b PA0 4"-deoxy-4"-epi-(phenylalanylamino)avermectin B1a/B1b PA0 4"-deoxy-4"-epi-(N-methoxycarbonyl-N-methylamino)avermectin B1a/B1b PA0 4"-deoxy-4"-epi-methoxycarbonylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-(2-methylpropyloxy)carbonylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-epi-phenoxycarbonylaminoavermectin B1a/B1b PA0 4"-deoxy-4"-(methylaminocarbonyl)aminoavermectin B1a/B1b PA0 4"-deoxy-4"-(phenylaminocarbonyl)aminoavermectin B1a/B1b PA0 4"-deoxy-4"-(2-propylaminocarbonyl)aminoavermectin B1a/B1b PA0 4"-deoxy-4"-(iminomethyleneamino)avermectin B1a/B1b PA0 4"-deoxy-4"-(dimethylaminomethyleneimino)avermectin B1a/B1b PA0 4"-deoxy-4"-(N-morpholinylmethyleneimino)avermectin B1a/B1b PA0 4"-epi-amino-4"-deoxyavermectin B1a/B1b N,N-dimethylacetamidine
There are eight different avermectin natural product compounds and they are given the designations A1a, A1b, A2a, A2b, B1a, B1b, B2a, and B2b based upon the structure of the individual compounds. In the foregoing structural formula, the individual avermectin compounds are as set forth below. (The R group is 4'-.alpha.(L-oleandrosyl).alpha.-L-oleandrosyloxy:
______________________________________ (A) R.sub.1 R.sub.2 R.sub.3 ______________________________________ A1a double bond -- sec-butyl --OCH.sub.3 A1b double bond -- iso-propyl --OCH.sub.3 A2a single bond --OH sec-butyl --OCH.sub.3 A2b single bond --OH iso-propyl --OCH.sub.3 B1a double bond -- sec-butyl --OH B1b double bond -- iso-propyl --OH B2a single bond --OH sec-butyl --OH B2b single bond --OH iso-propyl --OH ______________________________________
The avermectin compounds are generally isolated as mixtures of a and b components. Such compounds differ only in the nature of the R.sub.2 substituent and the minor structural differences have been found to have very little effect on the isolation procedures, chemical reactivity and biological activity of such compounds.
In addition to these natural avermectins containing the 25-iso-propyl or 25-sec-butyl-substituent, closely related derivatives containing other branched or cyclic 25-alkyl or 25-alkenyl substituents, optionally further substituted by heteroatoms such as oxygen, sulfur, nitrogen, and halogen, are known in the literature. These derivatives are obtained through various adjustments and additions to the fermentation procedures as described fully in the European Patent Application EPO 0 214 731.
Avermectins are products of microbial fermentations using the actinomycete Streptomyces avermitilis. These microbes use acetates and propionates as building blocks for most of the avermectin carbon chain, which is then further modified by microbial enzymes to give the completed avermectin molecules. It is known, however, that the carbon C-25 and the 2-propyl and 2-butyl substituents at this carbon are not derived from acetate or propionate units, but are derived from aminoacids L-valine and L-isoleucine, respectively. It was reasoned, that these aminoacids are deaminated to the corresponding 2-ketoacids, and that these then are decarboxylated to give 2-methylpropionic and 2-methylbutyric acids. These acids then have been found to be directly incorporated into the avermectin structures to give the 2-propyl and 2-butyl C-25 substituents, as is reported by Chen et al., Abstr. Pap. Am. Chem. Soc. (186 Meet.,MBTD 28, 1983). It was also disclosed in European Patent Application number 0 214 731 that additions of large amounts of other acids such as cyclopentanoic, cyclobutyric, 2-methylpentanoic, 2-methylhexanoic, thiophene-3-carboxylic acids and others to the fermentation broth of S. avermitilis causes the microbes to accept these acids as substitutes and to make small amounts of avermectins containing these acids in form of new C-25 substituents. Examples of such new avermectin derivatives are: 25-(thien-3-yl)-25-de-(1-methylpropyl)avermectin A2a 25-(cyclohex-3-enyl)-25-de-(1-methylpropyl)avermectin A2a 25-cyclohexyl-25-de-(1-methylpropyl)avermectin A2a 25-(1-methylthioethyl)-25-de-(1-methylpropyl) avermectin A2a 25-(2-methylcyclopropyl)-25-de-(1-methylpropyl) avermectin A2a
Similar experiments producing avermectins "c" and "d" containing as C-25 substituents a 2-pentyl and 2-hexyl group are described by T. S. Chen et al. in Arch. Biochem. Biophys. 1989, 269, 544-547.
Still additional avermectin derivatives are produced through artificial modification of the fermentation of Streptomyces avermitilis either by addition of metabolic inhibitors such as sinefungin (as described by Schulman et al., J. Antibiot. 1985, 38, 1494-1498) or by mutation of the parent strain (as described by Schulman et al., Antimicrobial Agents and Chemotherapy, 1987, 31, 744-747, and by EP-276-131-A to Pfizer INC.). Some of these avermectin derivatives are still further modified and are missing one or two of the 3'- and 3"-O-methyl groups (Schulman et al., J. Antibiot. 1985, 38, 1494-1498). Examples for such derivatives are:
The fermentation products have been chemically modified in order to obtain further antiparasitic and insecticidal analogs with improved properties. Publications of such procedures in the scientific and patent literature have been reviewed by Fisher, M. H.; Mrozik, H. In Macrolide Antibiotics; Omura, S., Ed.; Academic: New York, 1984; pp 553-606, and by Davies, H. G.; Green, R. H. Nat. Prod. Rep., 1986, 3, 87-121.
For example a group of semisynthetic avermectin derivatives were obtained by hydrogenating specifically the 22,23-double bond of avermectin B1 giving 22,23-dihydroavermectin B1 derivatives which have very potent anthelmintic and antiparasitic properties. Other examples of semisynthetic avermectin derivatives contain a 8,9-oxide group, a 4a-hydroxy or acyloxy group, a 23-keto group, which all are potent antiparasitic and insecticidal compounds.
It has also been described by Mrozik in U.S. Pat. No. 4,427,663 that amino substituents at the 4"- and 4'- positions have very high antiparasitic and insecticidal activities.
These compounds may be used as starting materials for the compounds of the instant invention without further modification, or when containing additional reactive groups, which are not to be modified under the reaction conditions applied, only after protection of such with a suitable protecting group.